JPS61122568A - Test composition for measuring glucose in high-concentrationregion made to be contained in aqueous test sample in semi-determination manner, testing tool and preparation of testing - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of the Invention The present invention relates generally to diagnostic reagent compositions containing enzymes, and more particularly to diagnostic reagent compositions containing enzymes in high concentration regions (h) contained in aqueous liquids.
(high range) enzymatic semi-quantitative measurement of glucose. [Usefulness] Semi-quantitative measurement of glucose contained in body fluids such as urine and blood is not only important as a means of mass screening for diabetes, but is especially important for diabetic patients who must control their sugar intake. It is. Because early diagnosis and continued control are so important in diabetes, glucose testing is extremely valuable to physicians, clinical departments, and those on diabetic diets at home. , must be quick and easy to perform, and at the same time be sensitive enough to detect significant changes in glucose in urine or blood;
The urine glucose concentration of diabetic patients is 5,000a+
g/dfL or more, so a degree 1,
0001g/di1. It is important to semi-quantitatively measure the urine glucose concentration in the high concentration region.
~ IQ, defined as glucose at a concentration of 000 mg/dJL or higher, and semi-quantitative evaluation of glucose in this high concentration range is useful for distinguishing and diagnosing diabetic coma from coma induced by drugs, alcohol, or trauma. At the very least, it is useful for therapeutic monitoring of insulin requirements.
It is important for two reasons. If the glucose concentration is very high when tested in an emergency vehicle, diabetic coma is suspected. Furthermore, if the dose of insulin is insufficient, the urine glucose level will be high, so a test that can semi-quantitatively measure glucose in the high concentration range is
It is also useful for therapeutic monitoring of insulin requirements. [Significance of the Invention] Clinically performed diagnostic glucose tests often rely on the enzymatic action of glucose oxidase on β-D-glucose [reaction formula (1)] and visually detected by a color change. Possible chromogen (Cr) oxidized state (
Cr t ) [Based on reaction formula (2) J] Peroxidase H2O2+Cr [20+ Cr
(2) This test uses a test device that can semi-quantitatively measure the glucose concentration by visually comparing the color developed after contact with the test sample using an appropriate color chart. Very convenient if possible. Such semi-quantitative measurements can also be performed by measuring the reflectance of the reacted test device with an instrument. U.S. Patent No. 2,848,308 and U.S. Patent No. 2,912,3
Previous patents, such as No. 09, all disclose glucose test "sticks," and the glucose test in these patents involves a microorganism containing glucose oxidase, peroxidase, a-tridine dihydrochloride, and boric acid. A stick coated with gelatin solution was dipped into a powder mixture. Approximately 75% by weight of this mixture is boric acid;
Boric acid is used as fillers such as talc, starch, sodium citrate-citric acid mixture, titanium oxide, and silica gel.
It is said that it can be replaced with. The glucose stick thus produced turned blue when moistened with a liquid containing glucose. Since then, various changes have been made to the components of glucose course test compositions, including enzymes, in order to improve techniques for semi-quantitatively identifying differences in glucose concentrations. However, even with many improved systems, when glucose concentrations exceed approximately 500 g/dIL, the color of the chromogen becomes too dark to distinguish between glucose concentration differences in high concentration regions. It is possible. GB Patent Specification No. 1.484.359 uses 0-tolidine, tetramethylbenzidine and tetraethylbenzidine as chromogens, and 0.50, 100. 25G,
The results of testing aqueous test samples containing 500 and 1,000 g/dl of glucose are described. And when the glucose concentration increases from 0 to 50 g/di, each of these chromogens changes color from yellow to bright green. However, the glucose concentration was 5006/
Beyond dl, the color of the oxidized chromogen becomes darker, and olive black, black, and dark green are observed for each chromogen, respectively. These results indicate that there are problems with semi-quantitative measurement of glucose at high concentrations, i.e., known chromogens produce a black or dark green color, so when measuring glucose exceeding 500 mg/di, such a test device may be used. This highlights the problem that its usefulness is diminished. Although such problems are less serious when color changes are measured using instruments, they still exist. In U.S. Pat. No. 4,340.11189, a visually measurable glucose concentration range is determined by using a glucose test device containing an enzyme and adding 1 liter of anisidine as a second chromogen. It has had some success in expanding. Moderate concentration region (i.e. 500
1 g/d to 1,000 g/dl), water-soluble iodide salts and poly(vinylpyrrolidone) are used as disclosed in U.S. Pat. No. 4,303,753. Glucose oxidase complex formation method using as a chromogen
Appropriate quantification can be performed using the following formulation. On the other hand, apart from the above, chelation of sugars with boric acid or its derivatives has been reported [Niss, A. Barker et al., [Carbohydrate Research].
Research) 28J 1973, pp. 33-40], However, to the best of the applicant's knowledge, this phenomenon has been used to solve problems in the measurement of glucose in the high concentration range contained in aqueous test samples. has not yet been used. [Summary of the Invention] The present invention provides a test composition, a test device, and a method for semi-quantitatively measuring glucose in a high concentration range contained in an aqueous test sample. an oxide active ingredient, a chromogenic indicator system capable of giving a detectable colorimetric response and p)! As used herein, high range glucose is characterized by comprising a borate buffer capable of providing an initial pH of greater than about 7. ,000 intestinal g/dfL ~ about 10,000 g
/d is defined as the glucose in the concentration range of the sentence. The test composition can also be integrated with a carrier matrix to form a solid state test device. Glucose in the high concentration region is
The present invention is particularly useful for visual semi-quantitative measurement of glucose in high concentration regions, which can be measured visually or instrumentally within 5 minutes, preferably about 2 minutes. DETAILED DESCRIPTION OF THE INVENTION Glucose oxidase test compositions comprising a borate buffer capable of providing an initial pH greater than about pH 7 are capable of semi-quantitatively measuring glucose in the high concentration range contained in an aqueous test sample. It turned out to be useful for measurements. A particularly preferred test format can be made by combining the test composition and a carrier matrix to form a solid state test device.Preferred applications of the invention include blood, serum, plasma, urine, etc. Although body fluids such as fluids, it should be understood that the test compositions, test devices and methods disclosed herein are also applicable to aqueous methods in the industrial field. In an experimental effort aimed at improving the discrimination of glucose concentration differences in the high concentration region, hexacitrate, phosphate, N,N'-bis(2-hydroxyethyl)
A number of buffers have been used with glucose oxidase/peroxidase systems, including glycine and N-2-hydroxyethylpiperazine, but significant improvements have been realized in distinguishing between glucose concentration differences in the high concentration region. is an initial pH greater than approximately pH 7.
This is only possible when the buffer is a boric acid group that can give . 1. Test Components Glucose oxidase test compositions as disclosed in U.S. Pat. No. 4,309,753 contain
In place of the buffer conditions hitherto used in such prior art, suitable semi-quantitative measurements of glucose at concentrations of 0.000 mg/di can be achieved, boric acid can provide an initial pH greater than about pH 7. By using a salt buffer, glucose in the high concentration range, i.e. about 1
,0001 g/di to about 10,000-g/di was significantly improved. In the present invention, a mixture of acid and base in the form Z-B(OH)2 is defined as a borate buffer. The equilibrium in such a borate buffer is approximately OH Z-B(OH)2+ 820 : Z-'9B
−OH+ H”OH [wherein the Z group may be a hydroxyl group (-OH) or some electron-withdrawing substituent]. For example, if Z is a hydroxyl group, the compound is represented by boric acid [
8(Q)I)3]. If 2 is a phenyl group, it becomes phenylporonic acid. Suitable poron dihydroxytes include boric acid, phenylporonic acid, p-nitrophenylporonic acid, 4-methoxyphenylporonic acid and other poronic acids and their derivatives.
In the present invention, the arene group is defined as a hydrocarbon containing at least one aromatic ring, and is an example of an electron-withdrawing group that can be used as a Z substituent, and the aromatic ring is 10 (OH
) The two anionic forms are positioned in such a way that they are stabilized by electronic resonance. Heterolene derivatives such as p-nitrophenylporonic acid containing an electron-withdrawing group as a substituent on the aromatic ring are also useful in the present invention. Borate buffers are 7-benboronic acid derivatives such as boric acid (Z-OH) and phenylporonic acid or Z-B(O
H) can be prepared from a mixture of two compounds by commonly used laboratory methods known to those skilled in the art. For example, a solution of borate buffer can be prepared by titrating boric acid with a base such as sodium hydroxide or potassium hydroxide to an initial pH of greater than pH 7. When using a borate buffer, the initial p)I is approximately 8
．． For use in dry test compositions, preferably between 5 and about 8.5, the water may be removed from the buffer, in which case the desired initial Powder it to obtain the desired pH. Suitable color indicator systems for use in the present invention include a combination of poly(vinylpyrrolidone) and a water-soluble iodide salt and 4-7 minoantipyrine with a suitable coupler, i.e., dichlorohydroxybenzenesulfonic acid, 4- Methylcatechol, 2,4-dihydroxybenzoic acid and 3
, 6-hydroxynaphthalene-2,7-disulfonic acid, etc. Poly(vinylpyrrolidone) is G
PVP-[1] from AF (New York, New York)
5 (average molecular weight 10,000), PVP-30 (
PVP-KftO (average molecular weight 40,000), PVP-KftO (average molecular weight 180,000), and PVP-Ni90 (average molecular weight 3 [tO,000)] are available. A preferred indicator system is a combination of poly(vinylpyrrolidone) and a water-soluble iodide salt;
Poly(vinylpyrrolidone) has a large molecular weight (PV
Preferred water-soluble iodide salts, most preferably P-90), include barium iodide, potassium iodide, sodium iodide, ammonium iodide and tetraalkylammonium iodide. The iodide salt must be capable of ionization, and the cationic portion of the salt must not interfere with the enzyme catalysis of glucose oxidation. Particularly preferred is potassium chloride. Chi Gua L, D -X 1' Gu -t/(E・0・
1, 1, 3, 4) #1 “Irus Volatories, Inc. (Elkhart, Indiana) or Sigma Chemical Company (
St. Louis, Missouri)
In the present invention, useful substances having peroxide activity are:
You can choose from a variety of organic and inorganic materials. For example, plant peroxidases can be used, such as horseradish or potato peroxidase. Although it is still not enough. Hemin, hemin derivatives, hemoglobin and hematin can also be used. Additionally, wetting agents, stabilizers, thickeners, etc. can be added, provided they do not interfere with the generation of a detectable response. A suitable wetting agent is available under the trade name Sarkosyl (5arkos2) from Sigma Chemical Co., St. Louis, MO.
Trauroylsarcosine available under 1@);
Trademark pending Emulphor ON 870 for polyoxyethylated eryl alcohol (ael!1alcohol) sold by GAF Inc. (New York, NY) and polyethylene glycol also available from Sigma Chemical Company. Triton X-100 is a registered trademark of Rohm & Hass Co., Ltd. for -p-yneoctylphenyl ether. The use of these components to make test devices is well known and therefore the selection of components does not require special skill. The test composition can be provided in the form of a bottled reagent, a frangible capsule containing the test composition in reagent form, a pill or a tablet. 2. Carrier Matrix A preferred form of the invention is made by treating a suitable carrier matrix with a test composition in the form of an aqueous reagent mixture. The carrier matrix is any material that can be incorporated with the components of the test composition as long as it is substantially inert with respect to the test composition and porous and/or absorbent with respect to the aqueous sample being tested. Any substance may be used. A "carrier matrix" is a hygroscopic or non-hygroscopic matrix that does not dissolve or have any structural change when exposed to water or other physiological fluids. Suitable hygroscopic matrices that can be used include paper, cellulose, wood, synthetic resin fleece, woven or non-woven fabrics. Suitable non-hygroscopic matrices that can be used include glass fibers, polymeric films, microporous membranes, and the like. Therefore, it should be recognized that all such carrier matrix concepts can be used in making the test devices of the present invention. In order to integrate the matrix and the constituent components, it is also possible to incorporate a system in which the components are uniformly combined in a liquid or semi-liquid state into the matrix, and then to cure the matrix, thereby integrating the components. Other matrix formats using porous membranes or polymer films are also contemplated. Microporous membranes can be prepared as ready-made membranes or by techniques such as phase inversion. Suitable polymer films can be made using commercially available latex formulations based on latex polymer suspensions formed by 80:40 copolymers of styrene and butadiene, for example. Other natural or synthetic polymers or mixtures thereof can also be used. An example of such a film formulation is U.S. Pat. No. 3,830,9
No. 57 and No. 4,312,834, which are only cited here as references. 3. Test device Preferably, the test device is prepared by integrating the carrier matrix and the test composition and drying them. In a preferred method of the invention, a hygroscopic carrier matrix (eg, filter paper) is impregnated with an aqueous solution of the test composition, dried, and the dried impregnated matrix is then applied to a support member. The solution with which the matrix is impregnated is prepared to obtain the desired initial DpH. When testing whole blood samples, dry impregnated carrier matrices can be coated with the sample and excess sample can be washed or scraped off in a manner that does not adversely affect the integrated test composition. Integration of the carrier matrix with the test composition may be accomplished by any means, preferably using an air oven, such as coating, dipping, spraying, spraying, or printing. This method can also be used. The dry carrier matrix can be cut and attached to one end of a support member, such as a rigid or semi-rigid polystyrene film strip. To immobilize the dry matrix on the support, use the 3M Company (St. Ball, Minn.) trademark Double Stick e.
Commercially available double-sided adhesive tape can be used. 4. Concentration range of test components The concentration range of the components in the reagent solution used to prepare the solid state test device is as follows. Possible range Preferred range Peroxy y Hze 200-5000 IJ/ml
1000-2000U/ml initial p
)I 7~10 8.5~
11.5 Instructions PVP-9G 0.1-5.0% 0.5-1.0
It is particularly preferred to use as high a concentration of borate as possible (i.e., 0.7 M to about 0.8 M) at an initial pH of about 9.0 to about 8.1%. used to integrate the test composition. Such borate buffers provide a favorable initial pH. That is, after drying the carrier, the surface of the test device is moistened and the buffer provides the desired initial pH. This surface pH can be measured using a surface electrode. The above concentration ranges and relative concentrations of the components are suitable for the aqueous polymer suspension used to incorporate the test components to make the polymer film, even if the reagent solution is an aqueous solution such as that used to impregnate a paper carrier matrix. It is effective even if it is a cloudy liquid. If a paper carrier matrix is used, a borate buffer, such as a phenylporonic acid buffer, is used to increase the pH to greater than about 7 prior to incorporating the test composition containing the borate buffer with the matrix. The paper can be pretreated at an initial pH. It is assumed that such pretreatment prevents interaction of the borate buffer in the test composition with the paper matrix. 5. How to use: The test device should be immersed in the test sample for a short time, or the test sample should be introduced into the carrier matrix. If present, a detectable colorimetric response develops. Contact with the sample can also be made using a pipette, swab or spartile. The immersion method is best when the sample is urine, but the use of a pipette is usually required when the sample is serum. Semi-quantitative glucose concentration measurements can be made by visually comparing the colors using a suitable color chart or by measuring the reflectance of the reacted test device with an instrument. If the support member is transparent,
Measurements can be taken from either side of the test device. The following examples are examples of experiments conducted in developing the present invention. However, these examples are provided to facilitate understanding of the present invention, and the present invention is not limited to these examples.
It should be possible to make changes and substitutions in the components and reaction parameters of the test composition that appear desirable. Abbreviations The following abbreviations are used in the following examples. 1 sentence
Milli lit 116 sentences
Deciliters g Otori Grams N Moles °C to Celsius POD Peroxidase GO Glucose Oxidase DH9A Dichlorohydroxybenzenesulfonic acid Poly (SiT-Go-MAA) Copolymer of styrene and methyl methacrylate FC&D Cloud 5 Yellow dye - Color Index Number 19140 PVP-90 Poly(vinylpyrrolidone) with an average molecular weight of 380,000 manufactured by GAF (=, - New York, York) Sarkosyl Surfactant N-lauroylsarcosine υ 1 International manufactured by Sigma Chemical Company (St. Louis, Missouri) Unit position (U) is the amount of enzyme that catalyzes the conversion of 1 micromole of substrate per minute. qs Amount of Solvent [Example] Example 1 A diborate buffer formulation Whatman 54IP paper was immersed in an aqueous solution containing the following ingredients. PVP- to 90G, 15
gmGO° 50,0000 POD 24,0000KI
O, 8g layer 5ark
os71 0. lsM borate buffer
14 layered water (IM, adjusted to pH 8.7 with sodium hydroxide) water
Remove excess solution with a qs 20mJl scraping rod and place the filter paper in an air oven at 50°C.
and dried for 20 minutes. After drying, double-sided adhesive tape was applied to the reagent paper and cut into ribbons with a width Q of 058 cm (115 inches). This ribbon was placed on one wide end of a long strip of polystyrene and cut into strips 0.058 cm (115 inches) wide to create test devices of the present invention. Glucose 0,500. 1,000.2,000
．． 9.000.5,000 and 10,000 mg/d
When the test device was immersed in an aqueous test sample containing Jl, a positive visual change was observed after 2 minutes of contact. Example 2: Comparison with an optimally prepared borate-free formulation A test device made according to Example 1 using a borate buffer was
An optimally prepared commercially available glucose oxidase/peroxidase test combination was compared with Diastix's (Miles Volatories, Inc., Ames Division, Elkhart, Ind.). Diastix formulation contains citric acid buffer (pH 7.2) and PVP/
Contains a KI indicator. For both formulations, test results were read at the time of maximum discrimination of the glucose concentration; this optimal reading time was 120 seconds for the borate formulation; For formulations without acid salts, it is 30 seconds. After these times, the color becomes darker for both formulations and the ability to distinguish concentrations decreases. To facilitate comparison between the two formulations, a Macbeth 1500 colorimeter was used with a Macbeth 1500 colorimeter. The reflectance was measured using the following method (manufactured by Kohlmorgen). 2 after contact with samples containing various concentrations of glucose
The ΔE between the test fixtures was calculated using these reflectance data. ΔE is a quantity indicating the overall color difference between two test devices in a three-dimensional color space. Calculated by the following formula [D. 8, Judd and G.
Colors in Business, Science and Industry (C
olor 1nBusiness, 5science
and Industry, ry) J. New York, see John Wiley and Sump (1975)]
. ΔE = [(ΔL books) 2 ◆ (Δa zero) 2 + (Δb books) 2]
'In the formula, ΔLt is an amount indicating the difference in brightness between two samples, and takes a value from O (complete black) to 100 (complete white). Δa volume is an amount indicating the difference between redness and greenness between two samples, and Δb is an amount indicating the difference between bright yellowness and blueness. 400-700nm for L-light, 6-wire, and B-wire.
Calculated from the reflectance of the wavelength. In the case of visual testing, the difference in density levels defined by the color chart must be large enough to distinguish the resulting color difference with the naked eye, and the difference between the two colors must be such that the ΔE between the two colors is If the chromaticity units are different, they can be identified with the naked eye. However, in practice, for one color to be distinguishable, the ΔE between different density levels must be 3 units or more; the larger the ΔE between density levels, the easier it is to distinguish the density levels with the naked eye. be. Citrate-PVP/Kl blend has glucose 1.0
When exposed to an aqueous test sample containing 0.00 mg/dJL, the color becomes greenish-yellow-brown and gradually becomes darker brown as the glucose concentration in the test sample increases; however, the borate-PVP/K I blend 50
When in contact with 0 g/d4 glucose, it is yellow;
As the glucose concentration increases, the brown color gradually becomes darker. ΔE between concentration levels Glucose concentration Citrate Borate negative 47...
・・・・・・・・・・・・251.000mg/dJ
L・・・・・・・・・・・・・・・・・・l・・・・・・
・・・・・・・・・・・・142.000mg/di ・・・・・・・・・・・・・・・3・・Fist・・・・
・・・・・・・・・・・・129.000-g/djL ・・・・・・・・・・・・・・・・・・2・・・・・・
・・・・・・・・・・・・13s, ooo intestine g/dl ・・・・・・・・・・・・・・・・・・ 1・・・・・・
・・・・・・・・・・・・1210.000 intestine g/d
Reading time: 30 seconds 120 seconds The value of ΔE between the concentration levels is determined by the glucose in the high concentration range (i.e., about 1,000 mg/di to about 10,000 tag
The results show that the borate formulation far outperforms the optimized commercial citrate formulation in measuring glucose in the range of /dfL). Example 3: Direct comparison where only the buffering agent was changed The borate formulations described above were compared with formulations where only the buffering agent was changed among the components or conditions. Borate blend Citric acid blend PVP-90 0.15gm
0.15gmGO50,000U
50,000 U, POD 24,0
00υ 24,000 UKl
G, 8g layer 0.6g
Amount 5arkosy+@0. *ml
O, 1i + boric acid 14 ta
n-(1%, pH with sodium hydroxide
Adjusted to 8,7) Citric acid/H2O-0, 85g water qs 20mIL
qs 20mfL pH 8,75,5 The final buffer concentration of each impregnation solution is 0.7M
Met. Glucose test devices were prepared in the same manner as described in Example 1 using each impregnating solution. The prepared test device was immersed in an aqueous solution containing a high concentration of glucose, and the concentration was measured using a Macbeth 1500 colorimeter (manufactured by Kohlmorgen, Newburgh, New York) to determine the concentration (1
The ΔE between the concentration and the concentration immediately above it was calculated. ΔE between concentration levels Glucose concentration Citrate Borate negative 51...
・・・・・・・・・・・・251.000mg/dj
L・・・・・・・・・・・・・・・・・・l・・・・・・
・・・・・・・・・・・・142.000mg/dJL ・・・・・・・・・・・・・・・＋13・・・・・・・・・
・・・・・・・・・・・・129.000 intestine g/dJL ・・・・・・・・・・・・・・・1・−・・
・・・・・・・・・・・・-135,000 intestine g/di ・・・・・・・・・・・・・・・0・・・・・・
・・・・・・・・・Hotland-121to, ooo■g/d
Sentence reading time 120 seconds 120 seconds The value of ΔE between concentration levels is the same for glucose in the high concentration region (i.e., glucose concentrations in the range of about t, ooo-g/d sentences to about to, oo. g/dl). In measurement,
It shows that the borate formulation is much superior to the citrate formulation. Example 4 = Comparison with conventional borate formulation Glucose oxidase/peroxidase formulation using boric acid as filler (U.S. Pat. No. 2,912,3
No. 09 and Example 2) of No. 2.848,308 were used as test compositions and impregnated into a carrier matrix. The thus prepared test device was tested for response to glucose in a high concentration range. P.O.D.
5 meals gGo
200 teng 80-tridine dihydrochloride
200mg boric acid 1800
Peng water 2
5tjL U.S. Patent Nos. 2,912,309 and 2
, 848,308 was dissolved in water to prepare an aqueous impregnating solution. This solution had a pH of 2.5, and the boric acid was not completely dissolved. Nevertheless, impregnation of Whatman 54 filter paper was carried out with this solution. The impregnated paper was dried at 50°C for 20 minutes. A test device was prepared from this impregnated paper as described above. The test device is negative, 1,000.2,000.
After 60 seconds of immersion in urine containing 4,000 and 8,000 sog/dfL of glucose, the colorimeter Macbeth 150
0 was used to record the value of ΔE (the longer the reading time, the darker the color and the smaller the value of ΔE). Glucose concentration ΔE Negative・・・・・・・・・・・・・・・・・・・・・・・・
OL, 000mg/d sentence・・・・φ・・・・・・・・・・・・・・・1
2.000 g/d sentence・・・・・・・・・・・・・・・・・・・・・1
4.000mg/d sentence・・・・・・・・・・・・・・・・・・・・・0
Therefore, the inability of conventional formulations using low pH boric acid as a "filler" to provide test compositions capable of distinguishing high concentration levels of glucose in aqueous test samples. it is obvious. Example 5: Pre-treated Paper Test Before combining with the composition, Whatman 5
4 Filter paper was soaked in an aqueous solution of 0.2M phenylporonic acid buffer (p
) 19.5). Let this pre-treated paper dry, then wait. Borate buffer (0.8M, pH 9.5) 5 t
anGO (12,500U/mJl)
0.8 tafLPOD (8,8000/mA
) 1. Om1DH9
A (0,5M)
1.0 m14-7 minoantipyrine (
IN) 0.25m! LFC & 5 yellow dye (1,88%) 0.3 so-called standing tree
qa 10. It was immersed in an impregnating solution containing Oto text. The test device prepared in this way has a concentration of 500 mg/dfL ~
responded to urine glucose concentrations in the range of 10.000 mg/dJL. A yellow dye is added to remove some strongly colored clinical floor disturbances. This dye is not a chromogen in the indicator system and has no effect on the visual identification of glucose concentration levels; the color of the test device changes from yellow (negative) to brown to red as the glucose concentration increases. 6 generates a detectable colorimetric response of approximately 45~B
Can be read in θ seconds, negative, 50G, 1
,000.2,000.9.000 and 5,000m
Visual discrimination of glucose in g/dJL is good. Example B: Polymer film from about 5005 g/dffi to about 10, in aqueous test sample
A test device that responds to 000 mg/diL glucose,
It can be made of a polymer film integrated with the test composition. One possible configuration is as follows. Part A 5 Potassium borate (IN, pH 9,5)
54 g Dispersant 0
．． 23 g Pigment 7,08
Layer antifoaming agent 0.007gm Surfactant 0.20 G layer thickener 〇J2 G layer part B Potassium borate (IM, pH 9,5) 4゜5 ■ Bun GO (5000U/a!L)
0.44sJLPOD (40
00U/m) 0.
90 for 4 m1PVP-
0.8 gyaKl
1. Og ■ Pa
- C Poly (ST-Go-MMA)
0.85gm dispersion (42% styrene) Par) Mix A and Part B and stir to form a homogeneous paste, then carefully add Part C with gentle stirring to prevent caking. The polymer suspension thus obtained is applied to a plastic support such as Tricite (polystyrene) and dried in an air oven at 50° C. for 20 minutes. The test device prepared in this way can be used in a high concentration range (i.e., 500 mg/dl to about 10.0 mg/dl).
f) can be reliably expected to respond to a range of urine glucose. Based on the technical idea of the present invention, it is clear that the present invention is not limited to the embodiments described above, and that various changes and modifications are possible.

Claims (1)

[Claims] 1. A test composition for semi-quantitatively measuring glucose in a high concentration range contained in an aqueous test sample, which comprises glucose oxidase, a peroxide active ingredient, and a detectable colorimeter. A color indicator system capable of giving a response and a pH of about 7
A test composition characterized in that it consists of a borate buffer that is capable of providing an initial pH that is greater than the initial pH. 2. The test composition according to claim 1, wherein the borate buffer is a borate buffer or a phenylboronic acid buffer. 3. The test composition according to claim 2, wherein the borate buffer is a borate buffer, and the color indicator system is a water-soluble iodide salt and poly(vinylpyrrolidone). 4. The test composition of claim 3, wherein said borate buffer is capable of providing an initial pH between about 8.5 and about 9.5. 5. A test device for semi-quantitatively measuring glucose in a high concentration range contained in an aqueous test sample, comprising: (a) a carrier matrix; (b) glucose oxidase, a peroxide active ingredient, and a detectable component; A color indicator system and p
A test device comprising a test composition integrated with said carrier matrix, comprising a borate buffer capable of providing an initial pH greater than about 7 H. 8. The test device according to claim 5, wherein the borate buffer is a borate buffer or a phenylboronic acid buffer. 7. The test device according to claim 6, wherein the borate buffer is a borate buffer, and the color indicator system is a water-soluble iodide salt and poly(vinylpyrrolidone). 8. The test device of claim 7, wherein said borate buffer is capable of providing an initial pH between about 8.5 and about 9.5. 9. A method for producing a test device for semi-quantitatively measuring glucose in a high concentration range contained in an aqueous test sample, the method comprising: (a) glucose oxidase, a peroxide active ingredient, giving a detectable response; Color indicator system and p
A test composition comprising about 0.2 molar to about 0.8 molar borate buffer capable of providing an initial pH greater than about H7 is combined with a carrier matrix and then (b) dried; A method characterized by comprising each step. 10. The method of claim 9, wherein the borate buffer is a borate buffer, and the color indicator system is a water-soluble iodide salt and poly(vinylpyrrolidone). 11. Claim 10, wherein the borate buffer is capable of providing an initial pH between about 8.5 and about 9.5.
The method described in section. 12. said carrier matrix is paper, and said method pre-treats said paper matrix with borate to an initial pH of about 7 prior to incorporation of said paper matrix with said test composition; 10. The method according to claim 9, comprising the step of: